KR102201231B1 - Pharmaceutical composition for preventing or treating gestational trophoblastic disease comprising butyl paraben - Google Patents

Abstract

The present invention relates to a composition comprising butyl paraben as an active ingredient, and more specifically, a pharmaceutical composition for preventing or treating gestational trophoblast disease comprising butyl paraben as an active ingredient and butyl paraben as an active ingredient It relates to a health functional food composition for preventing or improving gestational trophoblast disease.
The composition comprising butylparaben according to the present invention as an active ingredient is the proliferation of trophoblast cells by inhibiting phosphorylation of sub-signal transducers that regulate PI3K/AKT and MAPK signaling mechanisms in trophoblast cells that penetrate into the endometrium of the mother in early pregnancy. And the bar has the effect of inhibiting the permeability and improving the apoptosis effect, it can be usefully used in the field related to pharmaceuticals and functional foods capable of preventing and treating gestational trophoblast disease invented by abnormal development of trophoblast cells.

Description

Pharmaceutical composition for preventing or treating gestational trophoblastic disease comprising butyl paraben}

The present invention relates to a composition comprising butyl paraben as an active ingredient, and more specifically, a pharmaceutical composition for preventing or treating gestational trophoblast disease comprising butyl paraben as an active ingredient and butyl paraben as an active ingredient It relates to a health functional food composition for preventing or improving gestational trophoblast disease.

Gestational trophoblast diseases such as chorionic cancer and molar embryos are diseases caused by abnormal proliferation of trophoblast cells, excessive mobility and permeability. In particular, chorionic carcinoma is the most malignant disease among gestational trophoblastic diseases, and most of them develop from molar in early pregnancy (VCMak et al, Carcinogenesis, 2016; LDuffy et al, Journal of clinical medicine research, 2015), trophoblast tissue. Choriocarcinoma, a malignant tumor derived from, has been reported to appear at a rate of 1 per 20,000 to 40,000 people during pregnancy (Soper et al, Gynecol Oncol, 2004). In addition, choriocarcinoma is known to have very high vascular permeability and frequent metastasis to other organs such as lungs and vagina (Geramizadeh and Rad, Indian J Pathol Microbiol, 2012).

A common treatment for chorionic cancer is through chemotherapy called EMA-CO. However, 15-25% of patients with chorionic cancer show resistance to such chemotherapy and a poor prognosis (Powles et al, Br J Cancer, 2007). Therefore, there is a need for an approach through the development of more effective therapeutic agents in order to overcome the resistance to anticancer drugs in chorionic cancer.

Paraben is an alkyl ester compound contained in cosmetics, food, beverages, and pharmaceuticals and used as a preservative, has antimicrobial effects, and is known to regulate the endocrine system by mimicking the activity of estrogen (Zhang et al. , Toxicol Appl Pharmacol). , 2013). In particular, butylparaben has been reported to have stronger estrogen activity than other parabens such as methylparaben and ethylparaben (Daughton and Ternes, Environ Health Perspect, 1999).

As such, various pharmacological effects of butylparaben have been reported, but until now, nothing is known about the efficacy of butylparaben in relation to female reproductive or pregnancy-related diseases. In particular, the proliferation, mobility, and permeability of trophoblast cells There is no known effect on treating gestational trophoblast disease by modulating characteristics related to abnormal development.

Accordingly, the present inventors have made diligent efforts to develop a therapeutic agent for gestational trophoblast disease, as a result of confirming that butylparaben has the effect of inhibiting trophoblast cell proliferation, migration and permeability, and inducing apoptosis, and butylparaben has PI3K/AKT and ERK1 in trophoblast cells. The present invention was completed by confirming that it inhibits the /2 MAPK signaling mechanism.

An object of the present invention is to provide a pharmaceutical composition for preventing or treating gestational trophoblast disease comprising butylparaben as an active ingredient.

In addition, an object of the present invention is to provide a health functional food composition for preventing or improving gestational trophoblast disease comprising butyl paraben as an active ingredient.

In addition, an object of the present invention is to provide a method of inhibiting the proliferation, migration and penetration ability of trophoblast cells using the composition.

In addition, it is an object of the present invention to provide a method of improving the killing effect of trophoblast cells by using the composition.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating gestational trophoblast disease, comprising butylparaben as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving gestational trophoblast disease comprising butyl paraben as an active ingredient.

In addition, the present invention provides a method of inhibiting the proliferation, migration and penetration ability of trophoblast cells using the composition.

In addition, the present invention provides a method of improving the killing effect of trophoblast cells by using the composition.

The composition comprising butylparaben according to the present invention as an active ingredient is the proliferation of trophoblast cells by inhibiting phosphorylation of sub-signal transducers that regulate PI3K/AKT and MAPK signaling mechanisms in trophoblast cells that penetrate into the endometrium of the mother in early pregnancy. And the bar has the effect of inhibiting the permeability and improving the apoptosis effect, it can be usefully used in the field related to the pharmaceutical and functional food that can prevent and treat gestational trophoblast disease invented by abnormal development of trophoblast cells.

1 shows the results of analyzing the effect of butylparaben on the proliferation of trophoblast cell lines.
Figure 2 shows the results of analyzing the effect of butyl paraben on the apoptosis of trophoblast cell lines.
3 shows the results of analysis of active oxygen generation and calcium ion concentration change according to butylparaben treatment in trophoblast cells.
4 shows the results of measuring the mitochondrial membrane potential reduction effect according to the treatment of butylparaben in trophoblast cells.
5 shows the results of measuring the cell permeability inhibitory effect upon treatment of butylparaben on trophoblast cell lines.
6 shows the results of measuring the inhibitory effect of PI3K/AKT and ERK1/2 MAPK signaling pathways by butylparaben treatment in trophoblast cells.
7 shows the results of analyzing the phosphorylation pattern of signaling proteins according to the combination treatment of butylparaben, PI3K/AKT, and ERK1/2 inhibitor in trophoblast cells.
8 shows the mechanism of cell proliferation and cell death by butylparaben in trophoblast cells.

Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by an expert skilled in the art to which the present invention belongs. In general, the nomenclature used herein is It is well known and commonly used in the art.

In the present invention, the mechanism of cell death and inhibition of cell proliferation in trophoblast cells of butylparaben was revealed (FIG. 8).

In one aspect, the present invention relates to a pharmaceutical composition for preventing or treating gestational trophoblast disease comprising butylparaben as an active ingredient.

The present invention relates to a health functional food composition for preventing or improving gestational trophoblast disease comprising butyl paraben as an active ingredient from another perspective.

The term "composition" as used in the present invention is considered to include not only products containing specific ingredients, but also any products made directly or indirectly by the combination of specific ingredients.

In the present invention, the trophoblast cells may be cells derived from mammals, and the mammals are rodents (eg, mice, rats, hamsters, gerbils and guinea pigs), right heads (eg, cattle , Sheep, pigs, goats, deer, giraffes and antelopes), base titles (e.g. horses, donkeys, rhinos and tapirs), carnivores (e.g. dogs, cats, tigers, wolves, foxes, lions, cheetahs, Leopards, raccoons, badgers, cougars, jaguars and wildcats), order of rabbits (rabbits and crying rabbits), order of carnivores (such as hedgehogs, moles, and solenoidons) and primates (such as chimpanzees, orangutans, gorillas, Bonobono, Japanese monkey, rhesus monkey).

In the present invention, the pharmaceutical composition may be characterized in that it contains a pharmaceutically acceptable carrier, wherein the carrier is an ion exchange resin, alumina, aluminum stearate, lecithin, serum protein, buffer material, water, salt, Electrolyte, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substrate, polyethylene glycol, sodium carboxymethylcellulose, polyarylate, wax, polyethylene glycol and wool paper, characterized in that at least one selected from the group consisting of I can.

In the present invention, the pharmaceutical composition is formulated for intravenous, intraperitoneal, intramuscular, intraarterial, oral, intracardiac, intramedullary, intrathecal, transdermal, intestinal, subcutaneous, sublingual or topical administration. It may be characterized in that it further contains any one or more adjuvants selected from the group consisting of buffers, antimicrobial preservatives, surfactants, antioxidants, tonicity modifiers, preservatives, thickeners and viscosity modifiers, and solutions, suspensions, emulsions , It may be characterized by having a formulation selected from the group consisting of gels and powders.

A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as the severity of symptoms, weight, age, sex, mode of administration and time of administration of the patient, and generally skilled physicians are effective in the desired treatment or prevention. The dosage can be easily determined.

In the present invention, the health functional food refers to a food manufactured and processed using raw materials or ingredients having useful functions for the human body according to the Health Functional Food Act No. 6722, and contains nutrients for the structure and function of the human body. It refers to food consumed for the purpose of controlling or obtaining beneficial effects for health purposes such as physiological effects.

The health functional food composition of the present invention may be formulated into a formulation of a general health functional food known in the art, and granules, tablets, pills, suspensions, emulsions, syrups, gums, teas, jellies, various beverages, and drinks. , Alcoholic beverages, etc., and there is no particular limitation on the kind of the health functional food.

The health functional food composition of the present invention is in any herbal form suitable for administration to an animal body, including the human body, more specifically any form conventional for oral administration, for example, food or feed, additives and adjuvants for food or feed, It may be in a solid form such as fortified food or feed, tablets, pills, granules, capsules and foam formulations, or in liquid form such as solutions, suspensions, emulsions, beverages, pastes, etc., and nutrients, vitamins, electrolytes, sweeteners, colorants, organic acids, It may contain a preservative or the like, and these ingredients may be used independently or in combination.

According to the present invention, the butyl paraben may be characterized in that it inhibits PI3K/AKT and ERK1/2 MAPK signaling mechanisms.

According to the present invention, it may be characterized in that it further comprises a target inhibitor that inhibits the PI3K/AKT or ERK1/2 MAPK signaling pathway.

According to the present invention, the gestational trophoblast disease may be characterized in that it is at least one selected from the group consisting of choriocarcinoma, hydatidiform mole, and vilous adenoma.

The composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent.

In the present invention, it may be characterized in that it further comprises etoposide, cisplatin or paclitaxel, but is not limited thereto, and a chemotherapeutic agent used as a therapeutic agent for gestational trophoblast diseases including chorionic cancer, molar carcinoma, chorionic adenoma, etc. Can include.

In another aspect, the present invention relates to a method of inhibiting the proliferation, migration, and penetration ability of trophoblast cells by using the pharmaceutical composition.

In another aspect, the present invention relates to a method of improving the killing effect of trophoblast cells by using the pharmaceutical composition.

Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples. Therefore, it will be said that the practical scope of the present invention is defined by the appended claims and their equivalents.

Example

<Experiment method>

Experimental animals and cell culture

HTR8/SVneo cells were purchased from the American Type Culture Collection and used, and 5% fetal bovine serum (FBS) was mixed with RPMI-1640 medium containing 2.05 mM L-Glutaimine for monolayer culture. And used. For the experiment, monolayer cultured cells were cultured to a level of about 70% filling in a 100mm culture dish. Cells were starvated for 24 hours before treatment with butylparaben, and DMSO was used as a vehicle.

Experiment material

Candidate composition, butylparaben, was purchased from Selleckchem and used, and phospho-AKT, S6, P70S6K, GSK3β, ERK1/2, P90RSK protein and total-AKT, S6, P70S6K were used to confirm the signaling mechanism by butylparaben. , GSK3β, ERK1/2, P90RSK, IRE1α, antibodies against cytochrome C were purchased from Cell Signaling Techonology. In addition, antibodies against PCNA, GRP78, GADD153, ATF6α, and TUBA were purchased from Santa Cruz Biotechnology. LY294002 was purchased from Cell Signaling Technology, and U0126 from Enzon Life Science.

BrdU Cell proliferation ability analysis

To confirm the effect of butylparaben on the proliferation capacity of trophoblast cells, 5×10 ³ cells cultured under FBS starvation and 100 μL of medium were dispensed into 96 wells and dose-dependently (0, 50, 100, 200). , 400 μM) and cultured for 48 hours, and then an experiment was performed according to the manufacturer's manual using a BrdU kit (Cat No: 1167229001, Roche). After 48 hours of incubation, 10 μM BrdU was additionally added to each well and incubated in a 37° C./5% CO ₂ incubator for 2 hours. BrdU was labeled on choriocarcinoma cells, the cells were fixed, and the anti-BrdU-POD solution was incubated for 90 minutes at room temperature, followed by washing three times. Finally, the cells were washed with 100 μL of 3,3',5,5'-tetramethylbenzidine substrate. In response, the absorbance in 370 nm and 492 nm was measured using an ELISA reader to analyze the cell proliferation ability.

Immunofluorescence

After dispensing 3×10 ⁴ trophoblast cells in a confocal dish (catalog number: 100350, SPL Life Science, Republic of Korea) with 300 μl of 5% FBS-containing medium and incubating, an additional 24 hours FBS starvation After incubation, 200 μM of butylparaben was treated for 24 hours, then the cells were fixed with methanol for 10 minutes, and PCNA antibody diluted to 2 μg/ml was treated, and mouse IgG was treated as a control group and incubated at 4° C. for 16 hours. After that, after washing twice with PBS containing 0.1% BSA (bovine serum albumin), goat anti-mouse IgG Alexa 488 (catalog number: A-11001, Invitrogen, Carlsbad, CA, USA) was used as the secondary antibody. It was diluted 1:200 in antibody dilution buffer and incubated for 1 hour at room temperature. HTR8/SVneo cells were washed with 0.1% BSA-PBS, and then DAPI staining was additionally performed so that the nucleus as well as the target protein in HTR8/SVneo cells could be observed simultaneously. After the end of the experiment, the cells were observed and photographed using an LSM710 (Carl Zeiss, Thornwood, NY, USA) confocal microscope.

Annexin V and propidium Apoptosis analysis through iodide staining

In order to confirm the killing effect of trophoblast cells by butylparaben, an experiment was conducted using FITC Annexin V Cell Death Diagnosis Kit I (BD Biosciences). First, 5×10 ⁵ cells were cultured in 6 wells and further cultured in FBS starvation for 24 hours when the cells were filled in a 70-80% culture dish. Thereafter, butylparaben was dose-dependently treated (0, 50, 100, 200 μM) and cultured in a 37° C./5% CO ₂ incubator for 48 hours. Thereafter, the cells were removed from the culture dish using trypsin, washed with PBS, and the cells were slowly mixed and centrifuged using 1 mL of 1× binding buffer to obtain a cell pellet. Next, cell suspension culture was carried out with 200 μL of 1× binding buffer, 100 μL was added to a brown 1.5 mL tube, Annexin V 5 μL and PI 5 μL were mixed together, and the cells were left at room temperature for 15 minutes for staining. Then, 400 μL of 1× binding buffer was added, and the stained solution was transferred to a 5 mL FACS tube, and the fluorescence intensity was analyzed using a flow cytometer to measure the number of dead cells.

Protein expression analysis (Western blot)

After the trophoblast cells were treated with butylparaben, total protein was extracted from trophoblast cells, and the protein was quantified by Bradford protein assay (Bio-Rad, Hercules, CA, USA). Thereafter, the extracted protein was denatured at 95°C for 5 minutes, electrophoresis was performed using 10% SDS/PAGE gel, transferred to a nitrocellulose membrane, and the primary antibody and the secondary antibody were incubated sequentially, followed by chemiluminescence detection (SuperSignal West Pico, Pierce, Rockford, IL, USA) reagent was used to analyze the expression of the target protein using a ChemiDoc EQ system and Quantity One software (Bio-Rad) instrument.

Intracellular ROS measurement using DCFH-DA

To confirm the effect of butylparaben on ROS production in HTR8/SVneo cells, 2',7'-dichlorofluorescin diacetate (DCFH-DA, which is converted to 2', 7'-dichlorofluorescin (DCF) in the presence of peroxide and becomes fluorescent Sigma) was used. HTR8/SVneo cells were detached through trypsin and a cell pellet was obtained through centrifugation. After washing with PBS, 10 μM of DCFH-DA was incubated for 30 minutes in a 37°C incubator. The cells were then washed twice with PBS, and butylparaben was incubated in a 37°C incubator for 1 hour in a dose-dependent manner (0, 50, 100, 200 μM). The treated cells were washed again with PBS and analyzed for DCF fluorescence intensity using a flow cytometer.

Intracellular Calcium ion concentration measurement

In order to confirm the effect of butylparaben on the calcium ion concentration in HTR8/SVneo cells, 5×10 ⁵ cells were first cultured in 6 wells, and when the cells were filled in 70-80% culture dishes, they were further cultured in FBS starvation for 24 hours . Thereafter, butylparaben was treated in a dose-dependent manner (0, 50, 100, 200 μM) and incubated in a 37° C./5% CO ₂ incubator for 48 hours. Thereafter, cells were removed from the culture dish using trypsin, and Fluo-4 AM (Invitrogen) was incubated for 20 minutes in an incubator at 37°C. The stained cells were washed once with PBS and analyzed for fluorescence intensity using a flow cytometer.

JC Mitochondria through -1 staining Membrane potential Measure

JC-1 mitochondrial membrane potential (MMP) change was measured using a mitochondria staining kit (Cat No: CS0390, Sigma-Aldrich). 5×10 ⁵ trophoblast cells were cultured in 6 wells and further cultured in FBS starvation for 24 hours when the cells were filled in 70-80% culture dishes. Thereafter, butylparaben was dose-dependently treated (0, 50, 100, 200 μM) and cultured in a 37° C./5% CO ₂ incubator for 48 hours. Then, the cells were removed from the culture dish using trypsin and centrifuged to obtain a cell pellet. Cells were released in JC-1 staining solution and incubated for 20 minutes in a 37°C/5% CO2 incubator. The stained cells were centrifuged again, washed with 1x JC-1 staining buffer, and then analyzed for fluorescence intensity using a flow cytometer.

cell permeability analysis

In order to confirm the effect of butylparaben on the cellular invasiveness of trophoblast cells, Matrigel was coated on migration culture dishes and 200 μM of butylparaben was treated for 24 hours. Thereafter, the cells were fixed with methanol for 10 minutes, and then stained with hematoxylin solution (Sigma) for 30 minutes. The upper membrane, through which the cells did not pass, was washed with a cotton swab, and then mounted on a slide glass with a mounting medium (Sigma), and the cell penetration ability was observed, photographed and calculated with an optical microscope DM3000 (Leica).

Statistical analysis

The mean and standard error were calculated using SAS (statistical analysis system) statistical program, and one-way ANOVA was performed. A significance test was conducted at the P <0.05 level.

Results and Discussion

Trophoblast Cell proliferation Butyl paraben Impact Analysis

In order to analyze the change pattern of trophoblast cell line (HTR8/SVneo) in early pregnancy by butylparaben, it was first cultured for 48 hours in a medium added with butylparaben dose-dependently (0, 50, 100, 200, 400 μM), and then cell proliferation. As a result of analyzing the pattern, it was confirmed that the proliferation capacity of trophoblast cells was gradually reduced by butylparaben (FIG. 1A). The proliferation of trophoblast cells decreased by 42% ( P <0.001) in response to 100 μM butylparaben and 66% ( P <0.001) in response to 200 μM butylparaben. Then, immunofluorescence was performed using an antibody against PCNA, a protein essential for DNA proliferation (FIG. 1B). As a result, it was confirmed that the expression of PCNA was significantly reduced in the nuclei of trophoblast cells treated with 200 μM butylparaben for 24 hours compared to the control group. When this was quantified, butylparaben reduced the expression of PCNA by about 69% compared to the control. Through this, it was confirmed that butylparaben inhibits the proliferation of trophoblast cells.

Trophoblast Cell death Butyl paraben Impact Analysis

To analyze the apoptosis-inducing effect of butylparaben on trophoblast cells, apoptosis occurs through Annexin V and PI staining after 48 hours of incubation with butylparaben dose-dependent (0, 50, 100, 200 μM) on trophoblast cells. The number of trophoblast cells was measured using FACS. The cell group positive for Annexin V staining was assumed to be apoptosis.As a result, it was confirmed that cell death increased by 270% ( P <0.001) compared to the control when 200 μM of butylparaben was treated (FIG. 2A ). In addition, as a result of analyzing the expression of endoplasmic reticulum stress-related proteins such as GRP78, eIF2α, GADD153, ATF6α, and IRE1α, it was confirmed that the expression of each protein increased in a dose-dependent manner according to the administration of butyl paraben in trophoblast cells (Fig. 2B). In addition, Cytochrome C, a protein released from mitochondria as calcium ions increased, also increased in expression with butylparaben treatment. Through this result, it was confirmed that butylparaben induces the death of trophoblast cells, which is mediated by endoplasmic reticulum stress.

To butyl paraben by Trophoblast Analysis of mechanisms for regulating oxidative stress in cells

In order to confirm whether oxidative stress is induced in HTR8/SVneo cells by butylparaben, the degree of intracellular active oxygen (ROS) generation was confirmed through DCF fluorescence observation. DCF fluorescence increases with intracellular hydrogen peroxide generation. As a result, it was confirmed that the DCF fluorescence in HTR8/SVneo cells treated with butylparaben increased dose-dependently (0, 50, 100, 200 μM) (FIG. 3A). DCF fluorescence increased about 5 times by the highest concentration of 200 μM of butylparaben ( P <0.001). It is known that the increase of intracellular free radicals promotes the release of calcium ions from the endoplasmic reticulum and induces cell death. Therefore, fluo-4 staining was performed to analyze the change in intracellular calcium ion concentration, and as a result, it was confirmed that the intracellular calcium ion concentration increased in a dose-dependent manner by butylparaben (Fig. 3B). 200 μM of butylparaben increased the intracellular calcium ion concentration by about 2.1 times ( P <0.001) compared to the control. These results suggest that butylparaben induces an increase in free radicals leading to an increase in intracellular calcium ion concentration.

Morphological change of intracellular mitochondria is a representative mechanism of cell death caused by oxidative stress. JC-1 staining was performed to analyze whether the intracellular mitochondrial membrane potential changes according to the induction of intracellular oxidative stress by butylparaben. It was shown that the intracellular mitochondrial membrane potential decreased in a dose-dependent manner according to the administration of butylparaben in trophoblast cells (FIG. 4). It was found that 200 μM of butylparaben reduced the mitochondrial membrane potential by about 17.1 times ( P <0.001) compared to the control. Through this result, it was confirmed that butylparaben induces oxidative stress in cells, which also affects the mitochondrial membrane potential.

Trophoblast Intracellular Butyl paraben Cells according to treatment permeability Change analysis

It is essential for normal placental development that trophoblast cells have adequate permeability. To check the effect of butylparaben on the permeability of trophoblast cells, HTR8/SVneo cells were dispensed into Transwell coated with Matrigel, and incubated for 12 hours in a medium containing butylparaben (200 μM). As a result, the permeability of trophoblast cells was reduced by about 51% by butylparaben (FIG. 5A). In addition, it was measured how fast the trophoblast cells moved to fill the space between the two areas through the ibidi plate, and the mobility of the trophoblast cells treated with butylparaben was significantly lower than that of the control group (FIG. 5B). This suggests that butylparaben can inhibit excessive penetration of trophoblast cells.

PI3K / With AKT MAPK In the signaling pathway Butyl paraben Impact Analysis

In order to identify the signaling mechanisms that influence cell proliferation and apoptosis induced by butylparaben in trophoblast cells, the phosphorylation patterns of ERK1/2 MAPK and AKT related to proliferation were evaluated by butylparaben and PI3K/AKT specific inhibitors ( LY294002) and ERK1/2-specific inhibitor (U0126) were treated in combination and analyzed using Western blot (Fig. 6). As a result, it was confirmed that butylparaben not only inhibited phosphorylation of proteins included in the PI3K/AKT signaling pathway, such as AKT, P70S6K, GSK3β, S6, etc., but also increased the activity of ERK1/2 protein in trophoblast cells. The expression of P90RSK was not significantly affected by butylparaben. In addition, inhibition of PI3K/AKT and ERK1/2 signaling pathways affects each other's signaling pathways, suggesting that the signaling pathways regulated by butylparaben exhibit complex crossover effects. Treatment with 20 μM of LY294002 further reduced the phosphorylation of P70S6K and S6 reduced by butylparaben, and treatment with U0126 of 20 μM increased the phosphorylation of AKT and P70S6K inhibited by butylparaben.

In addition, the combination treatment of LY294002 or U0126 with butylparaben showed a result of further enhancing the antiproliferative effect and the apoptosis induction effect by butylparaben (FIGS. 7A, 7B). This effect was found to be further reduced by treatment with LY294002 or U0126 in combination with trophoblastic membrane potential (FIG. 7C). Cell proliferation was reduced by 26% ( P <0.001) by treatment with butylparaben and LY294002 and 39% by treatment with butylparaben and U0126 ( P <0.001). In addition, in terms of apoptosis, the combined treatment with LY294002 and U0126 increased by about 5.1 and 7.1 times ( P <0.001), respectively, compared to the treatment with butylparaben alone. In addition, the combination treatment of LY294002 and U0126 with butylparaben increased mitochondrial membrane potential reduction by about 3.8 and 221.3 times ( P <0.001), respectively, compared to butylparaben alone. Compared to the treatment with LY294002 alone, the addition of butylparaben further reduced cell proliferation, increased apoptosis and decreased mitochondrial membrane potential. These results suggest that the PI3K/AKT signaling pathway regulated by butylparaben is important in the change of physiological properties of trophoblast cells.

As described above, specific parts of the present invention have been described in detail, and it is obvious that these specific techniques are only preferred embodiments for those of ordinary skill in the art, and the scope of the present invention is not limited thereby. something to do. Therefore, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (7)

Contains butyl paraben as an active ingredient,
A pharmaceutical composition for preventing or treating gestational trophobastic disease selected from the group consisting of hydatidiform mole and villous adenoma. The method of claim 1,
The pharmaceutical composition for preventing or treating gestational trophobastic disease, characterized in that the butylparaben inhibits PI3K/AKT and ERK1/2 MAPK signaling mechanisms. The method of claim 1,
A pharmaceutical composition for preventing or treating gestational trophobastic disease, characterized in that it further comprises a target inhibitor that inhibits PI3K/AKT or ERK1/2 MAPK signaling pathway. delete Contains butyl paraben as an active ingredient,
Health functional food composition for preventing or improving gestational trophobastic disease selected from the group consisting of hydatidiform mole and villous adenoma. A method for inhibiting the proliferation, migration, and penetration ability of trophoblast cells derived from mammals other than humans by using a composition containing butyl paraben as an active ingredient. A method of improving the killing effect of trophoblast cells derived from mammals other than humans by using a composition containing butyl paraben as an active ingredient.

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